In the emerging field of digital pathology, images of tissue sample slides are digitally scanned and/or imaged, and saved as digital images. Each such image may consume as much as 10-to-25 Gigabytes (GB) of storage. The tissue sample, which is placed on the slide, may have a thickness of a few microns to a few millimeters. In some examples, the slides are scanned into a single two-dimensional (2D) image, where an autofocus algorithm determines and/or selects a focal plane for each area, region and/or portion of the image. In other examples, the slide is completely imaged and/or captured for and/or on each of a number of focal planes. Accordingly, a file representing the slide contains a plurality of 2D images for respective ones of a plurality of focal planes. The various 2D focal plane images can then be interpolated to allow a pathologist to interactively review any portion(s) of the digital slide at different focal planes.